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Extraction: Advanced Methods

Extraction: Advanced Methods

Metal ions can be separated from one another by complexation with organic ligands–the chelating agent– to form uncharged chelates. Here, the chelating agent must contain hydrophobic groups and behave as a weak acid, losing a proton to bind with the metal. Since most organic ligands used in this process are insoluble or undergo oxidation in the aqueous phase, the chelating agent is initially added to the organic phase and extracted into the aqueous phase. The metal-ligand complex is...

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Related Experiment Video

Updated: Dec 27, 2025

Biochemical Purification and Proteomic Characterization of Amyloid Fibril Cores from the Brain

Biochemical Purification and Proteomic Characterization of Amyloid Fibril Cores from the Brain

Published on: April 28, 2022

Amyloid-specific extraction using organic solvents.

Junichi Kamiie¹, Naoyuki Aihara¹, Yu Uchida¹

¹Laboratory of Veterinary Pathology, School of Veterinary Medicine, Azabu University, Sagamihara, Japan.

|March 7, 2020

Summary

Organic solvents can extract amyloid proteins from FFPE tissues, simplifying diagnosis. This new method bypasses laser microdissection for mass spectrometry-based amyloid typing.

Keywords:

Amyloid Amyloid-specific extraction from FFPE specimens using organic solvents Formalin-fixed Organic solvents Paraffin-embedded Protein extraction

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A Tailored HPLC Purification Protocol That Yields High-purity Amyloid Beta 42 and Amyloid Beta 40 Peptides, Capable of Oligomer Formation

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A Tailored HPLC Purification Protocol That Yields High-purity Amyloid Beta 42 and Amyloid Beta 40 Peptides, Capable of Oligomer Formation

A Tailored HPLC Purification Protocol That Yields High-purity Amyloid Beta 42 and Amyloid Beta 40 Peptides, Capable of Oligomer Formation

Published on: March 27, 2017

Area of Science:

Biochemistry
Proteomics
Histopathology

Background:

Amyloidosis diagnosis relies on mass spectrometry (MS) proteomic analysis.
Laser microdissection (LMD) is currently used for amyloid-specific sampling in MS analysis.
A simplified method for amyloid extraction from formalin-fixed, paraffin-embedded (FFPE) specimens is needed.

Purpose of the Study:

To establish a method for selectively extracting amyloids from FFPE specimens using organic solvents, eliminating the need for LMD.
To evaluate the efficacy of various organic solvents in amyloid extraction and characterization.

Main Methods:

Extraction of amyloids from FFPE mouse brain (Alzheimer's model) and human kidney (AA amyloidosis patient) using dimethyl sulfoxide (DMSO), dimethylformamide (DMF), methanol, trifluoroethanol (TFE), and hexafluoro-2-propanol (HFIP).
Analysis of extracts using SDS-PAGE, Western blotting, mass spectrometry, and circular dichroism spectrometry.
Comparison of organic solvent extraction with traditional LMD for amyloid typing.

Main Results:

Organic solvents (DMSO, DMF, methanol, TFE, HFIP) successfully extracted amyloid-beta (Aβ) from FFPE Alzheimer's mouse brain.
DMSO extracted serum amyloid A (SAA) from FFPE AA amyloidosis patient kidney tissue.
Circular dichroism revealed that TFE converted secondary structures of Aβ and transthyretin from β-sheets to α-helices.

Conclusions:

Organic solvents can effectively extract amyloids from FFPE specimens by altering their secondary structure.
This solvent-based extraction method simplifies amyloid typing by MS analysis and eliminates the LMD step.
The findings offer a more accessible approach for amyloidosis diagnosis and research.